LOCUS       sequence-363650-        7760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a Atm-targeting gRNA and Cre-recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7760)
  AUTHORS   Cai H, Chew SK, Li C, Tsai MK, Andrejka L, Murray CW, Hughes NW, 
            Shuldiner EG, Ashkin EL, Tang R, Hung KL, Chen LC, Lee SYC, Yousefi 
            M, Lin WY, Kunder CA, Cong L, McFarland CD, Petrov DA, Swanton C, 
            Winslow MM
  TITLE     A functional taxonomy of tumor suppression in oncogenic KRAS-driven 
            lung cancer.
  JOURNAL   Cancer Discov. 2021 Feb 19. pii: 2159-8290.CD-20-1325. doi: 
            10.1158/2159-8290.CD-20-1325.
  PUBMED    33608386
REFERENCE   2  (bases 1 to 7760)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cancer 
            Discov. 2021 Feb 19. pii: 2159-8290.CD-20-1325. doi: 
            10.1158/2159-8290.CD-20-1325."
COMMENT     Sequence Label: Lenti-sgAtm#1/Cre
FEATURES             Location/Qualifiers
     source          1..7760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(96..115)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        287..666
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        668..866
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     821..841
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             884..1064
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1111..1236
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1733..1966
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2151..2195
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2493..2610
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2692..2940
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2692..2712
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2863..2882
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2962..3037
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3090..3589
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3125..3148)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3522..3541
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3593..3613
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3614..4642
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     misc_feature    4698..5286
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4751..4771)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5169..5180)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5289..5305)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5290..5306)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5815..5995
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6057..6645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6137..6156)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6816..7607,7608..7676))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7439..7458
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7677..7760,1..21))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
       61 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
      121 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
      181 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
      241 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
      301 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      361 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      421 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      481 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      541 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      601 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      661 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      721 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      781 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      841 gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga
      901 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
      961 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     1021 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
     1081 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
     1141 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
     1201 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
     1261 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
     1321 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
     1381 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
     1441 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
     1501 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
     1561 accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca
     1621 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1681 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1741 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1801 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1861 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1921 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1981 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     2041 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     2101 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     2161 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     2221 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     2281 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     2341 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     2401 gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa
     2461 ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
     2521 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     2581 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
     2641 gtttggttag taccgggccc gctctagccc tgcaggaact gtttaaactg tgagggccta
     2701 tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga gataattgga
     2761 attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa
     2821 tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc
     2881 gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg accaaacacc
     2941 gcttctgtct ggagctctgt ggttttagag ctagaaatag caagttaaaa taaggctagt
     3001 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttgtttggc gcgcctatgg
     3061 cttaattaac atggcggccg cattctaccg ggtaggggag gcgcttttcc caaggcagtc
     3121 tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc
     3181 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt
     3241 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc
     3301 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac
     3361 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt
     3421 cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct
     3481 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa
     3541 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggacccggga tgcccaagaa
     3601 gaagaggaag gtgtccaatt tactgaccgt acaccaaaat ttgcctgcat taccggtcga
     3661 tgcaacgagt gatgaggttc gcaagaacct gatggacatg ttcagggatc gccaggcgtt
     3721 ttctgagcat acctggaaaa tgcttctgtc cgtttgccgg tcgtgggcgg catggtgcaa
     3781 gttgaataac cggaaatggt ttcccgcaga acctgaagat gttcgcgatt atcttctata
     3841 tcttcaggcg cgcggtctgg cagtaaaaac tatccagcaa catttgggcc agctaaacat
     3901 gcttcatcgt cggtccgggc tgccacgacc aagtgacagc aatgctgttt cactggttat
     3961 gcggcggatc cgaaaagaaa acgttgatgc cggtgaacgt gcaaaacagg ctctagcgtt
     4021 cgaacgcact gatttcgacc aggttcgttc actcatggaa aatagcgatc gctgccagga
     4081 tatacgtaat ctggcatttc tggggattgc ttataacacc ctgttacgta tagccgaaat
     4141 tgccaggatc agggttaaag atatctcacg tactgacggt gggagaatgt taatccatat
     4201 tggcagaacg aaaacgctgg ttagcaccgc aggtgtagag aaggcactta gcctgggggt
     4261 aactaaactg gtcgagcgat ggatttccgt ctctggtgta gctgatgatc cgaataacta
     4321 cctgttttgc cgggtcagaa aaaatggtgt tgccgcgcca tctgccacca gccagctatc
     4381 aactcgcgcc ctggaaggga tttttgaagc aactcatcga ttgatttacg gcgctaagga
     4441 tgactctggt cagagatacc tggcctggtc tggacacagt gcccgtgtcg gagccgcgcg
     4501 agatatggcc cgcgctggag tttcaatacc ggagatcatg caagctggtg gctggaccaa
     4561 tgtaaatatt gtcatgaact atatccgtaa cctggatagt gaaacagggg caatggtgcg
     4621 cctgctggaa gatggcgatt agccatgttt aagggttccg gttccactag gtacaattcg
     4681 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     4741 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     4801 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     4861 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     4921 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     4981 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     5041 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     5101 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     5161 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     5221 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     5281 ccccgcatcg ataccgtcga cctcgatcga gacctagaaa aacatggagc aatcacaagt
     5341 agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag
     5401 gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta
     5461 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     5521 agacaagata tccttgatct gtggatctac cacacacaag gctacttccc tgattggcag
     5581 aactacacac cagggccagg gatcagatat ccactgacct ttggatggtg ctacaagcta
     5641 gtaccagttg agcaagagaa ggtagaagaa gccaatgaag gagagaacac ccgcttgtta
     5701 caccctgtga gcctgcatgg gatggatgac ccggagagag aagtattaga gtggaggttt
     5761 gacagccgcc tagcatttca tcacatggcc cgagagctgc atccggactg tactgggtct
     5821 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     5881 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     5941 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagcatg
     6001 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6061 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6121 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6181 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6241 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6301 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6361 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6421 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6481 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6541 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6601 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6661 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6721 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6781 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6841 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6901 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6961 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7021 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7081 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7141 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7201 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7261 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7321 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7381 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7441 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7501 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7561 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7621 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7681 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7741 tttgaatgta tttagaaaaa
//
